低渗肿胀实验、苯胺蓝染色及染色质扩散实验检测精子核蛋白及膜完整性对比研究

Comparison of HOST,aniline blue staining test and chromatin diffusion test for judging the nucleoprotein and membrane integrity of sperms

目的通过对低渗肿胀实验(HOST)、苯胺蓝(AB)染色实验、染色质扩散(SCD)实验、脱氧核糖核酸末端转移酶介导的缺口末端标记(TUNEL)实验、AB/HOST实验,SCD/HOST实验、HOST/TUNEL实验的检测结果中精子活性分级进行比较,探索优化精子活性的检验方法。方法选取2012年5月—2016年10月已育成年健康男性志愿者30例。每例样本分为7组,分别为:HOST组、AB组、SCD组、TUNEL组、HOST/AB组和HOST/SCD组、HOST/TUNEL组。分别进行相应方法的染色,观察精子尾部肿胀及核蛋白、DNA受损情况,计算各级活力精子的比例。结果单独AB和单独HOST与HOST/AB比较,精子分级差异有统计学意义;单独SCD和单独HOST与HOST/SCD比较,精子分级差异有统计学意义;单独TUNEL和单独HOST与HOST/TUNEL比较,精子分级差异有统计学意义。结论 HOST/AB联合染色及HOST/SCD联合染色是对精子的核蛋白、DNA及膜完整性判断更为准确和实用的方法。

Objective Compare the sperm motility classification of hypoxia（ HOST）,aniline blue（ AB）staining,chromatin diffusion（ SCD）, the terminal deoxynucleotidyl transferase-mediated dU TP nick end labeling（ TUNEL） assay,HOST/AB,HOST/SCD and HOST/TUNEL test,to optimize testing method of sperm motility.Methods 30 healthy male volunteers were enrolled from May 2012 to October 2016. Each sample was divided into seven groups： HOST group,AB group,SCD group,TUNEL group,HOST/AB group,HOST/SCD group and HOST/TUNEL group. Respectively,after stainedin the corresponding method,observe the tail swelling,nuclear protein and DNA damage of the sperm,calculated theproportion of viable spermatozoa at all levels. Results There was significant difference of the classification of sperm between HOST,AB alone and HOST/AB test（ P 〈0. 01）. There was significant difference of the classification of sperm between HOST,SCD alone and HOST/SCD test（ P 〈0. 01）. There was significant difference of the classification of sperm between HOST,TUNEL alone and HOST/TUNEL test（ P〈 0. 01）. Conclusion HOST/AB,HOST/SCD and HOST/TUNEL test are more accurate and practical methods for judging the nucleoprotein,DNA and membrane integrity of sperms.